Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
  • G01N31/22—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using chemical indicators
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/483—Physical analysis of biological material
  • G01N33/487—Physical analysis of biological material of liquid biological material
  • G01N33/493—Physical analysis of biological material of liquid biological material urine
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10S436/00—Chemistry: analytical and immunological testing
  • Y10S436/807—Apparatus included in process claim, e.g. physical support structures
  • Y10S436/808—Automated or kit
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/20—Oxygen containing
  • Y10T436/200833—Carbonyl, ether, aldehyde or ketone containing
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
  • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
  • Y10T436/00—Chemistry: analytical and immunological testing
  • Y10T436/20—Oxygen containing
  • Y10T436/200833—Carbonyl, ether, aldehyde or ketone containing
  • Y10T436/202499—Formaldehyde or acetone

Definitions

• the present invention relates generally to the detection of aldehydes in such media as aqueous solutions, blood, urine and other body fluids and more specifically, to a reagent, the method of making the reagent, and a test kit for using the reagent in the detection of aldehydes and more specifically, malondialdehydes.
• the oxidative stress state can be measured from the release of aldehydes, particularly as dialdehydes, from the breakdown of long-chain polyunsaturated fatty acids.
• a fuchsin based calorimetric test can measure the released aldehyde in a rapid, easily performed home test kit by using the reagent described below with a small quantity of urine. The resulting color formed is compared to a calibrated test strip to assess the level of stress from a negative value through levels of +1 to +3.
• the first above-described test for example, requires heating of a test tube of solution in a constant temperature water bath for ten minutes. As a result of aldehyde tests not being quickly and easily performed, there may be a tendency for the tests for aldehydes not to be performed as frequently as they should or otherwise would be tested.
• An object of the invention is to provide a simple test for determining the presence of aldehyde in a test specimen.
• Another object is to provide a test reagent which produces a color change when mixed with a test specimen containing aldehyde.
• a testing solution or reagent for detecting the presence of aldehyde, particularly in an aqueous solution, urine and other body fluids comprises a solution of phosphoric acid, sodium metabisulfite, basic fuchsin and deionized water.
• the preferred proportions of the elements for a 1 liter batch are 10 grams sodium metabisulfite, 10 ml concentrated phosphoric acid, 5 grams basic fuchsin and about 1000 ml of water.
• Basic fuchsin changes color in an acidic solution, relative to the amount of aldehyde present in the specimen being tested.
• Basic fuchsin is a purple powder which reacts with aldehydes in the skin, urine or blood plasma. With low or no aldehydes present, you get no color development. With moderate or high levels of aldehydes you get color gradations roughly dependent on the level of aldehydes present.
• the amino group of the fuchsin couples with the aldehyde to produce the pink to purple color approximately dependent on the amount of aldehyde present in the biological fluids such as blood or urine. The color developed depends on the pH, which is controlled by the amount of acid present.
• Sodium metabisulfite is a reducer to stop the interference of oxygen from air. Metabisulfite ties up free oxygen so that only the aldehydes react with the fuchsin group. Establishing a nitrogen blanket over the reagent mixture gives greater shelf life to the reagent by stopping any oxygen reaction with the reagent. The phosphoric acid stabilizes the pH.
• Aldehydes are released from the breakdown of long chain polyunsaturated fatty acids by free radical attacks.
• High levels of aldehydes are found in a variety of diseases and abnormal metabolism states such as coronary artery disease, diabetes, and Parkinson disease.
• a method for testing for aldehydes in an aqueous solution, urine, blood and other body fluids comprises mixing a test specimen such as urine suspected of containing traces of aldehydes into the testing reagent described above, and observing for color formation after at least 2 minutes, preferably 2-5 minutes.
• the preferred proportions are about 0.9 to 1.1 ml of the test specimen to about 0.1-0.2 ml of the testing reagent. If the mixture does not become colored within such time, the test specimen contains less than about 2 ppm of aldehyde.
• the mixture becomes colored, for example, pinkish-purple, within the time specified, or sooner, aldehyde presence of greater than 2 ppm in the test specimen is inferred, the more intense the color, the greater the concentration of aldehyde.
• the testing reagent is contained in a sealed ampule or vial.
• Test specimen is introduced to the reagent in a snap-type ampule.
• a simple, efficient, reliable and rapid test for the presence of aldehyde in aqueous solution is thereby provided.
• the ampule along with an appropriate dispenser such as a syringe or pipette and a container for collecting the specimen are housed in a package bearing a color chart. This provides an easy, reliable way for an individual to test for the presence of aldehydes in his or her urine.
• a method of making the reagent is as follows: First, dissolve about 8-12 grams of sodium metabisulfite in about 995-1005 ml of dionized water, then add 10 ml of phosphoric acid and about 4.8 to 5.2 grams of basic fuchsin.
• the basic fuchsin is identified as BA 130 from Spectrum Chemical.
• the mixture should be mixed for at least 10 minutes and then add about 28-32 grams of animal bone charcoal.
• the mixture should then be well mixed and should remain standing for at least 24 hours and up to 36 hours. Thereafter, the bone charcoal is removed by centrifuging and filtering the mixture.
• the animal bone charcoal removes the color from the mixture thus, providing a decolorized solution.
• This decolorized solution is then adjusted for a pH between 1.75 and 1.93 by slowly adding phosphoric acid to the mixture and mixing it well during this procedure.
• a preferred method of making the reagent is as follows: First, dissolve 10 grams of sodium metabisulfite in 1000 ml of deionized water. Then add 10 ml of phosphoric acid and 5 grams of basic fuchsin (BA 130). The mixture should be mixed for at least 10 minutes. Then add 30 grams of animal bone charcoal. The mixture should then be well mixed and should remain standing for at least 24 hours. At this time, the bone charcoal is removed by centrifuging and filtering the mixture.
• the decolorized solution should be adjusted for pH.
• the pH should be gradually adjusted by adding phosphoric acid until it reaches a level of 1.88. It is important that the phosphoric acid be added slowly and mixed well during this procedure.
• the testing solution described above is preferably stored in individual, sealed test-size ampules or vials of conventional medical solution type. When packaged in such a manner and stored in a cool, dry place, the sealed bottle or vials have an expected shelf storage life of at least 12 months. Assurance of accurate testing solution may be achieved as descried below, by positive aldehyde test procedures.
• a test for the presence of aldehyde in an aqueous solution is then made by mixing about 0.9 ml of test solution (containing traces of aldehyde) into about 0.1 ml of testing solution formulated as above.
• the ampule is a 1 ml volume ampule containing 0.1 ml of the reagent sealed therein. The top of the ampule is broken off about a score line and discarded. The dispenser is used to fill the ampule with about 0.9 ml of urine. If the mixture of the test sample and testing solution remains colorless after a waiting period of about 2-5 minutes, the test is negative and the test sample therefore contains less than about 2 ppm aldehyde. Any color change of the mixture indicates presence of aldehyde in the test solution in a concentration greater than about 2 ppm.
• a positive aldehyde test is preferably by quality control techniques made before testing the test samples to assure that the testing solution is properly formulated or that, for example, the reagent bottles have not been replaced with other bottles containing non-testing solutions.
• the positive aldehyde test is preferably performed by injecting 1 ml of available “Positive Aldehyde Test Solution (Standard)” into a bottle containing about 0.2-0.6 ml of test solution. In approximately 2-5 minutes, the solution in the bottle should develop a pinkish-purple color provided the bottle contains properly formulated aldehyde testing solution. Otherwise, the bottle of “testing solution” from which the test bottle was selected should be discarded.
• the above-described positive test for aldehyde is sensitive to 10 ppm or more of aldehyde. For a 5 ppm, a positive test for aldehyde, 0.5 ml of deionized water is used. A colorless intense than that of the 10 ppm aldehyde test is obtained for the 5 ppm aldehyde test.

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• Health & Medical Sciences (AREA)
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• Urology & Nephrology (AREA)
• General Health & Medical Sciences (AREA)
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• Analytical Chemistry (AREA)
• Biophysics (AREA)
• Molecular Biology (AREA)
• Medicinal Chemistry (AREA)
• Food Science & Technology (AREA)
• Hematology (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
• Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

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• 2002
  • 2002-07-30 US US10/209,540 patent/US6689617B1/en not_active Expired - Lifetime
• 2003
  • 2003-07-29 EP EP03772087A patent/EP1525471B1/en not_active Expired - Lifetime
  • 2003-07-29 JP JP2004524194A patent/JP4354403B2/ja not_active Expired - Fee Related
  • 2003-07-29 DE DE60334003T patent/DE60334003D1/de not_active Expired - Lifetime
  • 2003-07-29 CN CNB038180367A patent/CN100338463C/zh not_active Expired - Fee Related
  • 2003-07-29 CA CA2494022A patent/CA2494022C/en not_active Expired - Fee Related
  • 2003-07-29 WO PCT/US2003/023811 patent/WO2004011910A2/en active Application Filing
  • 2003-07-29 KR KR1020057001788A patent/KR101034993B1/ko active IP Right Grant
  • 2003-07-29 AT AT03772087T patent/ATE479894T1/de not_active IP Right Cessation
  • 2003-07-29 AU AU2003259303A patent/AU2003259303A1/en not_active Abandoned
• 2005
  • 2005-08-11 HK HK05106905.6A patent/HK1073359A1/xx not_active IP Right Cessation

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